CRISPR-based genetic control strategies for insect pests

Genetic control strategies such as the sterile insect technique have successfully fought insect pests worldwide. The CRISPR(clustered regularly interspaced short palindromic repeats) technology, together with high-quality genomic resources obtained in more and more species, greatly facilitates the development of novel genetic control insect strains that can be used in area-wide and species-specific pest control programs. Here, we review the research progress towards state-of-art CRISPR-based genetic control strategies, including gene drive, sex ratio distortion, CRISPRengineered genetic sexing strains, and precision-guided sterile insect technique. These strategies’ working mechanisms,potential resistance development mechanisms, and regulations are illustrated and discussed. In addition, recent developments such as stacked and conditional systems are introduced. We envision that the advances in genetic technology will continue to be one of the driving forces for developing the next generation of pest control strategies.

CRISPR-mediated mutagenesis of the odorant receptor co-receptor(Orco)gene disrupts olfaction-mediated behaviors in Bactrocera dorsalis

Olfaction plays an essential role in insect behavior such as host location,foraging,mating,and oviposition.The odorant receptor co-receptor(Orco)is an obligatory odorant receptor and indispensable in odor perception.Here,we characterized the Orco gene from the oriental fruit fly,Bactrocera dorsalis(Hendel),a notorious agriculture pest.The olfactory deficiency mutants were generated by editing the BdorOrco gene using the CRISPR/Cas9 system.Electroantennograms(EAG)and olfactory preference assays confirmed that BdorOrco^(−/−)mutant flies had reduced perception of methyl eugenol,β-caryophyllene,and ethyl acetate.Oviposition bioassays showed that the eggs laid by BdorOrco^(−/−)females mediated by benzothiazole and 1-octen-3-ol were significantly decreased.In addition,BdorOrco^(−/−)mutant flies took a significantly longer time to locate the food source compared with wild type(WT)flies.Altogether,our data indicated that Orco is essential for multiple physiological processes in B.dorsalis,and it expands our understanding of the function of insect Orco.

High-Performance Liquid Chromatography Fingerprint of Marine Traditional Chinese Medicine Haliotidis

Objective: Haliotidis, the dried muscle of abalone, has long been used as a marine traditional Chinese medicine(TCM) due to its high medicinal and nutritive values, whereas few reports contributed to its quality control. This study aimed to establish the high-performance liquid chromatography(HPLC) fingerprints of Haliotidis. Materials and Methods: The HPLC fingerprints of Haliotidis, the manufacturing products of dried muscle of abalone, were analyzed by reverse-phase HPLC, using a YMC HPLC C18 column(250 mm × 4.6 mm, 5 μm).The gradient elution mode was applied with mobile phase starting from 5% acetonitrile and 0.1% phosphoric acid aqueous solution to 100%acetonitrile in 70 min at a flow rate of 1.0 mL/min. The detection wavelength was set at 190 nm. Results: Through compared the HPLC fingerprint chromatograms of 14 batches of Haliotidis, 25 peaks were assigned as the characteristic common peaks, of which one index component L-phenylalanine was identified by comparison with its standard substance. The established HPLC fingerprint analysis method of Haliotidis was validated with good precision, repeatability, and stability. Based on the above study, a reference HPLC fingerprint chromatogram of Haliotidis was proposed. Conclusions: It is the first time to establish the HPLC fingerprint chromatogram of TCM Haliotidis, which could contribute to the establishment of its quality control system.